scholarly journals High efficiency TALENs enable F0 functional analysis by targeted gene disruption in Xenopus laevis embryos

Biology Open ◽  
2013 ◽  
Vol 2 (5) ◽  
pp. 448-452 ◽  
Author(s):  
K.-i. T. Suzuki ◽  
Y. Isoyama ◽  
K. Kashiwagi ◽  
T. Sakuma ◽  
H. Ochiai ◽  
...  
Keyword(s):  
Functional Analysis ◽  
Xenopus Laevis ◽  
Gene Disruption ◽  
High Efficiency ◽  
Targeted Gene Disruption ◽  
Xenopus Laevis Embryos

scholarly journals Targeted gene disruption in Xenopus laevis using CRISPR/Cas9

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Fengqin Wang ◽  
Zhaoying Shi ◽  
Yan Cui ◽  
Xiaogang Guo ◽  
Yun-Bo Shi ◽  
...  
Keyword(s):  
Xenopus Laevis ◽  
Gene Disruption ◽  
Targeted Gene Disruption

scholarly journals Targeted Inactivation of Francisella tularensis Genes by Group II Introns

2008 ◽  
Vol 74 (9) ◽  
pp. 2619-2626 ◽  
Author(s):  
Stephen A. Rodriguez ◽  
Jieh-Juen Yu ◽  
Greg Davis ◽  
Bernard P. Arulanandam ◽  
Karl E. Klose
Keyword(s):  
Francisella Tularensis ◽  
Gene Disruption ◽  
Molecular Mechanisms ◽  
High Efficiency ◽  
Single Step ◽  
Group Ii Introns ◽  
Targeted Gene Disruption ◽  
Genetic Techniques ◽  
Group Ii ◽  
Targeted Inactivation

ABSTRACT Studies of the molecular mechanisms of pathogenesis of Francisella tularensis, the causative agent of tularemia, have been hampered by a lack of genetic techniques for rapid targeted gene disruption in the most virulent subspecies. Here we describe efficient targeted gene disruption in F. tularensis utilizing mobile group II introns (targetrons) specifically optimized for F. tularensis. Utilizing a targetron targeted to blaB, which encodes ampicillin resistance, we showed that the system works at high efficiency in three different subspecies: F. tularensis subsp. tularensis, F. tularensis subsp. holarctica, and “F. tularensis subsp. novicida.” A targetron was also utilized to inactivate F. tularensis subsp. holarctica iglC, a gene required for virulence. The iglC gene is located within the Francisella pathogenicity island (FPI), which has been duplicated in the most virulent subspecies. Importantly, the iglC targetron targeted both copies simultaneously, resulting in a strain mutated in both iglC genes in a single step. This system will help illuminate the contributions of specific genes, and especially those within the FPI, to the pathogenesis of this poorly studied organism.

scholarly journals Development of the CRISPR/Cas9 System for Targeted Gene Disruption in Aspergillus fumigatus

2015 ◽  
Vol 14 (11) ◽  
pp. 1073-1080 ◽  
Author(s):  
Kevin K. Fuller ◽  
Shan Chen ◽  
Jennifer J. Loros ◽  
Jay C. Dunlap
Keyword(s):  
Aspergillus Fumigatus ◽  
Gene Disruption ◽  
Polyketide Synthase ◽  
High Efficiency ◽  
Constitutive Expression ◽  
Targeted Mutagenesis ◽  
Genomic Alteration ◽  
Loss Of Function ◽  
Content Type ◽  
Targeted Gene Disruption

ABSTRACTLow rates of homologous recombination have broadly encumbered genetic studies in the fungal pathogenAspergillus fumigatus. The CRISPR/Cas9 system of bacteria has recently been developed for targeted mutagenesis of eukaryotic genomes with high efficiency and, importantly, through a mechanism independent of homologous repair machinery. As this new technology has not been developed for use inA. fumigatus, we sought to test its feasibility for targeted gene disruption in this organism. As a proof of principle, we first demonstrated that CRISPR/Cas9 can indeed be used for high-efficiency (25 to 53%) targeting of theA. fumigatuspolyketide synthase gene (pksP), as evidenced by the generation of colorless (albino) mutants harboring the expected genomic alteration. We further demonstrated that the constitutive expression of the Cas9 nuclease by itself is not deleterious toA. fumigatusgrowth or virulence, thus making the CRISPR system compatible with studies involved in pathogenesis. Taken together, these data demonstrate that CRISPR can be utilized for loss-of-function studies inA. fumigatusand has the potential to bolster the genetic toolbox for this important pathogen.

scholarly journals Iron nanoparticle bio-interactions evaluated in Xenopus laevis embryos, a model for studying the safety of ingested nanoparticles

2019 ◽  
Vol 14 (2) ◽  
pp. 196-213
Author(s):  
Patrizia Bonfanti ◽  
Anita Colombo ◽  
Melissa Saibene ◽  
Luisa Fiandra ◽  
Ilaria Armenia ◽  
...  
Keyword(s):  
Xenopus Laevis ◽  
Iron Nanoparticle ◽  
Xenopus Laevis Embryos ◽  
Ingested Nanoparticles

Teratogenic effects of five anticancer drugs on Xenopus laevis embryos

2016 ◽  
Vol 133 ◽  
pp. 90-96 ◽  
Author(s):  
Marina Isidori ◽  
Concetta Piscitelli ◽  
Chiara Russo ◽  
Marie Smutná ◽  
Luděk Bláha
Keyword(s):  
Xenopus Laevis ◽  
Anticancer Drugs ◽  
Teratogenic Effects ◽  
Xenopus Laevis Embryos
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